This study aimed to assess the initial multicenter experience using the robotic device — the iSR’obot Mona Lisa — to perform MRI-US fusion transperineal biopsy. The iSR’obot Mona Lisa performs robotic MRI-ultrasound fusion transperineal prostate biopsy via two skin punctures safely and with high accuracy. Overall, targeted cores detected at least as many significant cancers as template cores. Targeted and template biopsies together is most optimal for cancer grading and detection.